Nutritional composition

ABSTRACT

A nutritional composition is provided which has an effective prophylactic and/or improving effect for various symptoms of muscle mass decrease, decreased basal metabolism, low body temperature, suppression of obesity, suppression of visceral fat accumulation, hyperglycemia, hyperlipidemia and the like in elderly people. The nutritional composition can include n-3 fatty acid and one or more of free lysine, dipeptides containing lysine, and lysine salts, wherein the content of free lysine, dipeptides containing lysine, and/or lysine salts is 0.1 g-10.0 g per 100 kcal of the composition, and the content of the total amount of n-3 fatty acid is 0.17 g-5.00 g per 100 kcal of the composition.

This application is a Continuation of, and claims priority under 35U.S.C. §120 to, International Application No. PCT/JP2012/060186, filedApr. 13, 2012, and claims priority therethrough under 35 U.S.C. §119 toJapanese Patent Application No. 2011-089626, filed Apr. 13, 2011, theentireties of which are incorporated by reference herein. Also, theSequence Listing filed electronically herewith is hereby incorporated byreference (File name: 2013-10-11T_US-504_Seq_List; File size: 1 KB; Daterecorded: Oct. 11, 2013).

FIELD OF THE INVENTION

The present invention relates to a nutritional composition containingparticular amounts of a particular amino acid and n-3 fatty acid, whichis effective for increasing muscle mass and/or suppressing a decrease inthe muscle mass. More particularly, the present invention relates to anutritional composition containing particular amounts of one or more ofthe following: free lysine, dipeptides containing lysine and lysinesalts, and n-3 fatty acid, which is effective for increasing muscle massand/or suppressing a decrease in the muscle mass.

BRIEF DESCRIPTION OF THE RELATED ART

In elderly people, a specific skeletal muscle decrease called sarcopeniacan occur. A decrease in skeletal muscle reduces the amount of activityelderly people are able to do, and the decreased amount of activity cancause further loss of muscle strength, thus forming a vicious circle. Adecrease in the amount of activity also can decrease the basal energymetabolism. This condition is particularly apparent when in a bedriddenstate.

Moreover, elderly people have a high risk of falling whenundernourished. According to a search of the Social Insurance Institutein Japan, about 40% of hospital patients and about 30% of home-carepatients are undernourished. Even when energy ingestion is increased inan attempt to improve the nutrient condition, muscle mass cannot berecovered. Conversely, fat accumulates and leads tometabolism-associated diseases such as insulin resistance, diabetes,hyperlipidemia, and osteoporosis (Clinical Nutrition, 2008, 8: 38-43).

To counter muscle mass decrease, protein ingestion is recommended. Infact, it has been reported that ingestion of 18 g of an essential aminoacid mixture increased protein synthesis of the skeletal muscle (Am JClin Nutr, 2003, 78: 250-258). However, ingesting a large amount ofamino acids, such as up to 18 g, all at once not only is difficult forelderly people due to their depressed kidney function due to aging, butalso is risky since it may over burden on the kidneys. To increase themuscle mass in elderly people, therefore, not only supplementing with ageneral protein supplementation, but also ingestion of a more efficientcomposition is necessary.

Ingestion of lysine is known to increase growth and muscle mass incommunity residents and animals living on corn and wheat(JP-A-2008-539793). However, this is because the protein composition ofthe staple food is not balanced and lysine becomes a limiting aminoacid. In most areas where protein is not necessarily obtained largelyfrom corn and wheat, ingestion of lysine alone does not show a cleareffect on the growth of the whole body, or an increase in the musclemass that appears as one manifestation thereof.

JP-A-H11-508282 discloses a composition for patients with cachexiaand/or anorexia, which contains a mixed oil with a weight ratio of n-6fatty acid to n-3 fatty acid of 0.1-3.0, amino acid containing branchedchain amino acid, and antioxidant such as β-carotene, vitamin C, vitaminE, and selenium. In addition, Br J Cancer, 2004, 91: 408-412 reportsthat a carcinoma cachexia model mouse ingested with n-3 fatty acid and ahigh leucine-containing protein diet shows a decrease in the body weightand an increase in the muscle mass. In the target patients, particularcytokines (TNF-α, IL-6 and the like) due to cachexia in the late stagesof cancer are involved, and the muscle protein secretion decrease occursbased on the cytokines. It is assumed that n-3 fatty acid suppressessecretion of the cytokines, and branched chain amino acid such asleucine aids muscle protein synthesis, whereby the decrease of muscleproteins can be prevented. In other words, this composition was studiedfor treating carcinoma cachexia wherein cytokine-dominant proteindecrease was extreme. However, this decrease is vastly different fromthe muscle mass decrease in general elderly people experience due to adecreased amount of activity by being bedridden and the like, and theeffect is unknown.

Although n-3 fatty acid is used as a hypertriglyceridemia improvingagent, a single use of n-3 fatty acid has been reported to show noeffect on the weight increase of muscle (Drugs of the Future, 2010, 35:41-52 and J Nutr Biochem, 2006, 17: 1-13).

While n-3 fatty acid has been reported to increase mRNA expression of anenzyme involved in fat oxidation (J Nutr, 2005, 135: 2503-2508),enhanced energy metabolism causing enhanced synthesis of protein such asmuscle has not been confirmed to date.

As mentioned above, as the situation stands, a nutritional compositionhaving an appropriate amino acid ratio, containing n-3 fatty acid, andhaving a function to maintain or increase the muscle mass of elderlypeople has not been provided heretofore.

SUMMARY OF THE INVENTION

A novel nutritional composition is described having a preventive and/orimproving effect on a decrease in the muscle mass, particularly saideffect for elderly people with a decreased muscle mass due to adecreased amount of activity.

A composition is described that contains particular amount(s) of one ormore kinds of free lysine, dipeptides containing lysine and lysinesalts, and a particular amount of n-3 fatty acid that can prevent and/orimprove a decrease in the muscle mass, can improve hyperglycemia andhyperlipidemia by suppressing obesity and suppressing accumulation ofvisceral fat, and such effect is remarkable particularly in elderlypeople.

It is an aspect of the present invention to provide a nutritionalcomposition comprising n-3 fatty acid and an ingredient selected fromthe group consisting of free lysine, dipeptides containing lysine,lysine salts, and combinations thereof; wherein the ingredient ispresent in the composition in an amount of 0.1 g-10.0 g per 100 kcal ofthe composition, and wherein the n-3 fatty acid is present in thecomposition in an amount of 0.17 g-5.00 g per 100 kcal of thecomposition.

It is a further aspect of the present invention to provide thenutritional composition as described above, further comprising abranched chain amino acid selected from the group consisting of valine,leucine, and isoleucine.

It is a further aspect of the present invention to provide thenutritional composition as described above, wherein the branched chainamino acid is present in the composition in an amount of 1.0 g-20 g per100 kcal of the composition.

It is a further aspect of the present invention to provide thenutritional composition as described above, wherein the branched chainamino acid in a free form is present in the composition in an amount of0.1 g-15 g per 100 kcal of the composition.

It is a further aspect of the present invention to provide thenutritional composition as described above, wherein the n-3 fatty acidis selected from the group consisting of eicosapentaenoic acid,docosapentaenoic acid, docosahexaenoic acid, and combinations thereof.

It is a further aspect of the present invention to provide thenutritional composition as described above, comprising 10 mg-3000 mg ofeicosapentaenoic acid per 100 kcal of the composition.

It is a further aspect of the present invention to provide thenutritional composition as described above, further comprising a vitaminselected from the group consisting of:

0.2 mg-20.0 mg of vitamin B1 per 100 kcal of the composition,

0.25 mg-20.0 mg of vitamin B2 per 100 kcal of the composition,

0.3 mg-20.0 mg of vitamin B6 per 100 kcal of the composition,

0.3 μg-20.0 μg of vitamin B12 per 100 kcal of the composition, and

combinations thereof.

It is a further aspect of the present invention to provide thenutritional composition as described above, further comprising a vitaminselected from the group consisting of:

50 μg retinol equivalents-2500 μg retinol equivalents of vitamin A per100 kcal of the composition,

10 mg-700 mg of vitamin C per 100 kcal of the composition,

1 mg-50 mg of vitamin E based on α-tocopherol per 100 kcal of thecomposition, and

combinations thereof.

It is a further aspect of the present invention to provide thenutritional composition as described above, comprising 1 mg-50 mg ofzinc per 100 kcal of the composition and/or 1.0 μg-50.0 μg of seleniumper 100 kcal of the composition.

It is a further aspect of the present invention to provide thenutritional composition as described above, comprising medium chainfatty acid oil in an amount of 10 wt %-65 wt % relative to the totalamount of lipid.

It is a further aspect of the present invention to provide thenutritional composition as described above, which is capable ofincreasing muscle mass and/or suppressing a decrease in the muscle mass.

It is a further aspect of the present invention to provide thenutritional composition as described above, which is capable ofdecreasing visceral fat or increasing energy consumption.

It is a further aspect of the present invention to provide thenutritional composition as described above, which is capable of aneffect selected from the group consisting of prophylaxis and/orimprovement of muscle weakness symptom, promotion of rehabilitationeffect for muscle recovery, prophylaxis and/or improvement ofdyslipidemia associated with visceral fat increase, prophylaxis and/orimprovement of hyperglycemia associated with visceral fat increase,prophylaxis and/or improvement of fatty liver associated with visceralfat increase, prophylaxis and/or improvement of deteriorated liverfunction associated with visceral fat increase, prophylaxis and/orimprovement of decrease in basal energy consumption, prophylaxis and/orimprovement of decrease in basal body temperature, prophylaxis and/orimprovement of muscle mass decrease unaccompanied by an increase ininflammatory cytokine, improvement of low nutrient condition,improvement of deterioration in locomotorium, prophylaxis and/orimprovement of locomotive syndrome, prevention of falling, increase ofmuscle mass in sports, and combinations thereof.

It is a further aspect of the present invention to provide a method forincreasing muscle mass and/or suppressing a decrease in muscle mass,comprising administering an effective amount of the composition asdescribed above.

It is a further aspect of the present invention to provide a method fordecreasing visceral fat or increasing energy consumption, comprisingadministering an effective amount of the composition as described above.

It is a further aspect of the present invention to provide a method fortreating a condition comprising administering an effective amount of thecomposition as described above, wherein said condition is selected fromthe group consisting of prophylaxis and/or improvement of muscleweakness symptom, a method for the promotion of rehabilitation effectfor muscle recovery, a method for the prophylaxis and/or improvement ofdyslipidemia associated with visceral fat increase, a method for theprophylaxis and/or improvement of hyperglycemia associated with visceralfat increase, a method for the prophylaxis and/or improvement of fattyliver associated with visceral fat increase, a method for theprophylaxis and/or improvement of deteriorated liver function associatedwith visceral fat increase, a method for the prophylaxis and/orimprovement of decrease of basal energy consumption, a method for theprophylaxis and/or improvement of decrease of basal body temperature, amethod for the prophylaxis and/or improvement of muscle mass decreaseunaccompanied by an increase in inflammatory cytokine, a method for theimprovement of low nutrient condition, a method for the improvement ofdeterioration in locomotorium, a method for the prophylaxis and/orimprovement of locomotive syndrome, a method for prevention of falling,a method for increasing muscle mass in sports, and combinations thereof.

A method for producing an agent for increasing muscle mass and/orsuppressing a decrease in muscle mass comprising formulating acomposition as described above.

It is a further aspect of the present invention to provide a method forproducing an agent for decreasing visceral fat or increasing energyconsumption comprising formulating a composition as described above.

It is a further aspect of the present invention to provide a method forproducing an agent for treating a condition comprising formulating acomposition as described above, wherein the condition is selected fromthe group consisting of: the prophylaxis and/or improvement of muscleweakness symptom, promoting of rehabilitation effect for musclerecovery, the prophylaxis and/or improvement of dyslipidemia associatedwith visceral fat increase, the prophylaxis and/or improvement ofhyperglycemia associated with visceral fat increase, the prophylaxisand/or improvement of fatty liver associated with visceral fat increase,the prophylaxis and/or improvement of deteriorated liver functionassociated with visceral fat increase, the prophylaxis and/orimprovement of decrease of basal energy consumption, the prophylaxisand/or improvement of decrease of basal body temperature, theprophylaxis and/or improvement of muscle mass decrease unaccompanied byan increase in inflammatory cytokine, improvement of low nutrientcondition, improving deterioration in locomotorium, the prophylaxisand/or improvement of locomotive syndrome, prevention of falling,increasing muscle mass in sports, and combinations thereof.

The nutritional composition as described herein is effective forincreasing muscle mass and/or suppressing a decrease in muscle mass. Inaddition, the nutritional composition is effective for increasing energyconsumption. In addition, the nutritional composition is particularlyuseful for decreasing visceral fat based on the effect of increasingenergy consumption by enhanced fat utilization and the like. Inaddition, it is useful for the prophylaxis and/or improvement ofdyslipidemia, hyperglycemia, fatty liver, and deterioration of liverfunction associated with visceral fat increase. Furthermore, byincreasing muscle mass or suppressing a decrease in muscle mass by fatenergy supply and the like, the nutritional composition is also usefulfor the prophylaxis and/or improvement of sarcopenia, chronicobstructive pulmonary disease (COPD), promotion of rehabilitation effectfor muscle recovery, improvement of low nutrient condition, improvementof deterioration in locomotorium, prophylaxis and/or improvement oflocomotive syndrome, prevention of falling, increasing muscle mass insports and the like. Moreover, the nutritional composition is effectivefor the prophylaxis and/or improvement of a decrease in basal energyconsumption, and prophylaxis and/or improvement of a decrease in basalbody temperature. Furthermore, the nutritional composition is highlysafe, and can be used for a long time without causing an excessiveprotein load even for elderly people with attenuated kidney function.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows mesenteric fat weight and gastrocnemius muscle weight ofgroups 1A-1D of Experimental Example 1.

FIG. 2 shows mesenteric fat weight and gastrocnemius muscle weight ofgroups 2A-2D of Experimental Example 2.

FIG. 3 shows respiratory quotient and energy consumption of groups 3Aand 3B of Experimental Example 3.

FIG. 4 shows a ratio of DNA amount of mitochondria gene (mtDNA) to thatof nuclear coding gene (chDNA) of groups 3A and 3B of ExperimentalExample 3.

DESCRIPTION OF THE PREFERRED EMBODIMENTS

The nutritional composition can include one or more of free lysine,dipeptides containing lysine, and/or lysine salts in a total amount of0.1 g-10.0 g per 100 kcal of the composition, and n-3 fatty acid in atotal amount of 0.17 g-5.00 g per 100 kcal of the composition.

Lysine is one of the essential amino acids, and is the limiting aminoacid when corn, wheat, and the like are staple foods, and isparticularly effectively when added to the feed for domestic animals,since it contributes to the efficient growth of the animal. In addition,lysine may be in any form as long as it is in a free form, dipeptideform, salt form, or the like (hereinafter the “lysine” can mean aconcept encompassing free lysine, dipeptides containing lysine, lysinesalts and the like). Since the lysine in the nutritional composition isin a free form, a dipeptide form, a salt form or the like, thecomposition can be used for a long time without causing an excessiveprotein load, even for elderly people with attenuated kidney function.While the lysine may be in the L form, D form, or DL form, the L form isa particular example. The lysine in various forms can be used alone, ortwo or more forms of lysine can be used in combination. While the formof lysine is not limited, examples of the lysine salts include acidaddition salt, salt with base and the like. Examples of the acid to beadded to lysine to form a salt include inorganic acids such as hydrogenchloride, hydrogen bromide, sulfuric acid, phosphoric acid; organicacids such as acetic acid, lactic acid, citric acid, tartaric acid,maleic acid, fumaric acid, monomethylsulfuric acid; acidic amino acidssuch as glutamate, aspartic acid, and the like. In addition, examples ofthe base that forms a salt with lysine include hydroxide orcarbohydroxide of metal such as sodium, potassium, calcium, inorganicbases such as ammonia; organic bases such as ethylenediamine,propylenediamine, ethanolamine, monoalkylethanolamine,dialkylethanolamine, diethanolamine, and triethanolamine. More specificexamples of the lysine salts include lysine hydrochloride, lysineacetate, lysine glutamate, lysine aspartate and the like. “Dipeptide”refers to an amino acid dimer wherein two amino acid molecules arepeptide-bonded, and the “dipeptides containing lysine” can refer to adipeptide wherein at least one molecule, or 2 molecules, of thedipeptide is lysine. Examples of the dipeptides containing lysineinclude lysyllysine and the like. Among these lysines, lysine in a saltform can be used, and L-lysine hydrochloride, L-lysine acetate, andL-lysine glutamate are particular examples when used in food. Moreover,since flavor, pH upon dissolution, and the like can vary depending onthe form, the form of lysine can be appropriately changed depending onthe use.

The amount of lysine can be 0.1 g-10.0 g per 100 kcal of the compositionand, from the aspect of amino acid nutrition balance, can be 0.2-5.0 g,or 0.25-4.5 g. When the amount of lysine is less than 0.1 g per 100 kcalof the composition, the desired effect cannot be expected since thelysine is consumed making body protein or an energy source. Conversely,when the amount exceeds 10.0 g, a large amount of a single amino acid isingested, which is not very desirable for the amino acid balance. Theamount of lysine can be based on the free form.

While the daily intake of lysine is individually determined depending onthe age, sex, body weight, diet condition, and the like, it can be 20mg-400 mg, or 40 mg-200 mg, per kg human body weight, from the aspect ofamino acid nutrition balance. While the maximum tolerable intake oflysine per day has not been clarified, when it is more than 20 g perhuman kg body weight, a large amount of a single amino acid is ingested,which is not very desirable for the amino acid balance.

The n-3 fatty acid can be an unsaturated fatty acid having a double bondat the third position from the methyl group end of the hydrocarbonchain. Examples thereof include eicosapentaenoic acid, docosahexaenoicacid, α-linolenic acid, docosapentaenoic acid, and the like. These maybe used alone, or two or more kinds thereof may be used in combination.

The n-3 fatty acid is abundantly present in fats and oils such as fishoil, Japanese basil oil, and flaxseed oil. The n-3 fatty acids can beextracted from these fats and oils and used after purification. It isalso possible to use n-3 fatty acids produced by a chemical synthesismethod, a fermentation method and the like, and a commercially availableproduct for food can also be used. As a lipid source, moreover, the fatsand oils that contain an abundant amount of n-3 fatty acid can also beused directly.

The total amount of n-3 fatty acid present in the composition can be0.17 g-5.00 g, 0.18 g-4.00 g, or 0.19 g-3.00 g, per 100 kcal of thecomposition. When the total amount is less than 0.17 g per 100 kcal ofthe composition, a clear effect cannot be expected. Conversely, anamount exceeding 5.00 g in total per 100 kcal of the composition is notvery preferable in view of the flavor. In Japan, ingesting not less than2 g of n-3 fatty acid per day is recommended. Fats and oils containingn-3 fatty acid may be directly used, or n-3 fatty acid or fats and oilsthat contain an abundant amount of n-3 fatty acids may be mixed withother oils and used, as long as the amount of the n-3 fatty acid fallswithin the aforementioned range.

The n-3 fatty acid can contain eicosapentaenoic acid, docosapentaenoicacid, and/or docosahexaenoic acid. When the n-3 fatty acid containseicosapentaenoic acid, the amount of eicosapentaenoic acid can be 10mg-3000 mg, 20 mg-2000 mg, 50 mg-1000 mg, per 100 kcal of thecomposition. In addition, the amount of eicosapentaenoic acid relativeto the total amount of n-3 fatty acid can be 1 wt %-60 wt %, or 5 wt%-40 wt %. In Japan, ingesting not less than 1 g of eicosapentaenoicacid together with docosahexaenoic acid per day is recommended, and theupper limit is not particularly set.

The nutritional composition can further contain a total amount of 1.0g-20 g of one or more kinds of branched chain amino acids, such asvaline, leucine and/or isoleucine, per 100 kcal of the composition.

The branched chain amino acid can be valine, leucine, and/or isoleucine.The form of these branched chain amino acids is not particularly limitedand may be any form such as a protein form, a peptide form, a salt form,or a free form. In consideration of the burden on the kidney and in anattempt to not increase the overall amount of protein, the free form isusually the most preferable. In this case, the L form, D form, or DLform can be used. In addition, as the aforementioned branched chainamino acid, any of the amino acids obtained by extraction from animalsand plants etc., and purification thereof, and the amino acids obtainedby a chemical synthesis method, a fermentation method, or a generecombination method, may be used.

The above-mentioned branched chain amino acids have been found to bemore highly effective when present in not less than about 0.1 g in totalper 100 kcal of the composition, and an increased amount of the branchedchain amino acid can be used when a stronger effect is expected. Inconsideration of degradation of the taste, the amount thereof per 100kcal of the composition can be 1.0 g-20 g, or 1.2 g-12 g, in total. Whenthe total amount of the branched chain amino acids excesses 20 g per 100kcal of the composition, a large amount of a particular amino acid isingested, which is not very preferable for the amino acid balance.

The branched chain amino acid may be in any form such as a free form, apeptide form, a protein form, or salt form. The branched chain aminoacids may be all one type of form, or may be a mixture of two or morekinds of different forms. For example, the branched chain amino acid mayconsist solely of a branched chain amino acid in a free form, or maycontain a branched chain amino acid in a free form and a branched chainamino acid in a protein form, and the like. When the nutritionalcomposition contains a branched chain amino acid in a free form, thetotal amount thereof can be 0.1 g-15 g, or 0.25-12 g, per 100 kcal ofthe composition.

The amount of branched chain amino acid is typically expressed based onthe free form.

The branched chain amino acid can be valine, leucine, and/or isoleucine.In this case, the mixing ratio of isoleucine, leucine, and valine can begenerally isoleucine:leucine:valine=1.0:1.5-3.0:0.5-1.5 in a weightratio. Particularly, the ratio of leucine can be increased to about 1.5-to 3-fold relative to valine.

The nutritional composition can contain B vitamin.

Examples of B vitamins include vitamin B₁ such as thiamine; vitamin B₂such as riboflavin; vitamin B₆ such as pyridoxine, pyridoxal,pyridoxamine; vitamin B₁₂ such as cyanocobalamin. These may be usedalone or in combinations of two or more. B vitamins play an importantrole in the metabolism of carbohydrates, protein, and lipid. The amountof B vitamins can be 0.2 mg-20.0 mg for vitamin B₁, 0.25 mg-20.0 mg forvitamin B₂, 0.3 mg-20.0 mg for vitamin B₆, and 0.20 μg-10.0 μg forvitamin B₁₂, per 100 kcal of the composition.

In addition, the nutritional composition can also contain antioxidantvitamin and antioxidant mineral. Examples of the antioxidant vitamininclude vitamin A such as retinol, retinal, retinoic acid; vitamin Csuch as ascorbic acid; carotenoid such as β-carotene; vitamin E such asα-tocopherol, and the like. Examples of the antioxidant mineral includezinc, selenium, and the like. These may be used alone, or incombinations of two or more. When fat is utilized as an energy sourcefor muscle protein synthesis, since excess radicals are produced andoxidative stress is caused as a result, an antioxidant can be added forthe prevention and/or improvement thereof. To achieve sufficientantioxidant activity, the amount of antioxidant vitamin and antioxidantmineral can be 50 μg retinol equivalents-2500 μg retinol equivalents forvitamin A, 10 mg-700 mg for vitamin C, 1 mg-50 mg based on α-tocopherolfor vitamin E, 1 mg-50 mg for zinc, and 1.0 μg-50.0 μg for selenium, per100 kcal of the composition.

The nutritional composition can also contain vitamins other than Bvitamins and antioxidant vitamins (e.g., vitamin D, vitamin K, niacin,folic acid, pantothenic acid, biotin etc.), and minerals other thanantioxidant minerals (e.g., sodium, calcium, iron, phosphorus,magnesium, potassium, copper, iodine, manganese, chrome, molybdenumetc.). These may be used alone, or in combinations of two or more.

In addition, the nutritional composition can contain a medium chainfatty acid oil as a lipid source. The “medium chain fatty acid” can meana fatty acid having 8-10 carbons, such as caprylic acid, capric acid.The term “medium chain fatty acid oil” can also mean fats and oilsincluding triglycerides of the aforementioned medium chain fatty acid,and the like. The medium chain fatty acid can be characterized in thatit is digested and absorbed about 4 times more rapidly than long chainfatty acids that are generally present in fats and oils, delivered afterabsorption in the liver via the portal vein without passing throughlymphatic vessels, and rapidly metabolized. Therefore, it can bepreferably utilized as an energy source. Fats and oils containing alarge amount of medium chain fatty acids such as coconut oil, palm oil,palm kernel oil can be used as the medium chain fatty acid oil. Coconutand palm oils, and the like, can be extracted and purified from naturalplants such as coconut. However, a commercially available product can beconveniently used. Medium chain fatty acid oils can be added in anamount of 10 wt %-65 wt % relative to the total weight of lipid.

The nutritional composition can contain, as a fat source other thanmedium chain fatty acid oil, for example, edible vegetable oil such ascottonseed oil, sunflower oil, peanut oil, canola oil, soybean oil,safflower oil, olive oil, rice oil, corn oil, benne oil, cacao butter,edible animal oil such as beef fat, lard, fish oil, butter, butter oil,and processed fats and oils such as shortening. These may be used alone,or in combinations of two or more.

The nutritional composition can contain, as a carbohydrate source, forexample, glucide, dietary fiber, and the like. Examples of glucideinclude dextrin, oligosaccharide, saccharose, glucose, fructose, starch,and the like. Examples of dietary fiber include water-soluble dietaryfiber (e.g., indigestible dextrin, pectin, galactomannan etc.),insoluble dietary fiber (e.g., soybean- and wheat-derived bran,crystalline cellulose etc.), and the like. These may be used alone, orin combination of two or more kinds.

The nutritional composition can contain as a protein source, aminoacids, peptides, proteins, and the like. As the protein, animal-derivedprotein such as casein, acid casein, casein sodium, casein calcium, wheyprotein, milk serum whey protein, fish protein, egg protein, andhydrolysates thereof, and plant-derived protein such as soybean protein,wheat protein, corn protein, and hydrolysates thereof, and the like canbe added. The total amount of these protein sources in the nutritionalcomposition can be 2-20 g per 100 kcal of the composition. When theseprotein sources contain branched chain amino acids, one should includethe amount of the branched chain amino acid from these sources whencalculating the total amount of the branched chain amino acids in thecomposition.

When the nutritional composition contains a branched chain amino acid ina protein form, the total content of the protein source in thenutritional composition also including the protein can be 2-20 g per 100kcal of the composition. Particularly, when the chosen branched chainamino acid in the nutritional composition consists only of a branchedchain amino acid in the free form, the content of the total amount ofthe protein source in the nutritional composition can be 1.0-15 g per100 kcal of the composition.

The nutritional composition can be directly administered into the bowelsand stomach of patients showing insufficient oral ingestion, by using anadministration tube, or when oral ingestion is possible, it can be givenas a food or drink. Lysine and n-3 fatty acid, and further, one or morekinds of branched chain amino acids such as valine, leucine and/orisoleucine, and the like, may be directly mixed and ingested as anutritional composition, or the composition can also be formulated witha pharmaceutically acceptable carrier and provided as a pharmaceuticalproduct. Alternatively, the composition can also be provided in the formof food or drinks, such as food claiming specified health or nutritionalbenefits, by adding to a food or drink, or a food additive, or can beprovided as a general food or drink.

The nutritional composition can be formulated as a liquid preparationsuch as an elixir, suspension, syrup, emulsion, ampoule; or a solidpreparation such as gel, gum, drop, powder, granule, pill, tablet(including sugar-coated tablet, film-coated tablet), capsule, packageagent, powder, and the like.

Examples of the pharmaceutically acceptable carrier, which can be usedfor formulating the nutritional composition, include cellulose and aderivative thereof such as crystalline cellulose,hydroxypropylcellulose; excipients such as natural polymer compound (gumarabic, sodium alginate etc.); binders such as guar gum, stearic acid,polymeric polyvinylpyrrolidone; lubricants such as talc, polyethyleneglycol 6000; disintegrants such as adipic acid, surfactants such assucrose fatty acid ester, soybean lecithin, polyoxyethylene hydrogenatedcastor oil, polyoxyethylene monostearic acid ester; thickeners such assodium carboxymethylcellulose, carboxyvinyl polymer, xanthan gum,gelatin; coating agents such as ethyl acrylate-methyl methacrylatecopolymer dispersion, caramel, Carnauba wax, shellac-pullulan; pHadjusters such as citric acid, sodium citrate, acetic acid, sodiumacetate, sodium hydroxide; antioxidants such as erythorbic acid,butylhydroxyanisole, propyl gallate; flavoring agents such as aspartame,licorice extract, saccharin; preservatives such as sodium benzoate,sodium edetate, sorbic acid, sodium sorbate, methyl paraoxybenzoate,butyl paraoxybenzoate; colorants such as ferric oxide red, yellow ironoxide, black iron oxide, carmine, Food Color blue No. 1, Food Coloryellow No. 4, Food Color Red No. 2, and the like.

When the nutritional composition can be provided as food and drink, suchproducts can include liquid products such as drinks, milk products suchas milk, milk beverage, yogurt, jelly products such as jelly drinks,jelly, gum products, powder products, granular products, sheet products,capsule products, tablet products, solid products such as snack bar,cookie, and the like.

Examples of materials which can be used for forming the nutritionalcomposition as a food or drinks include a sweetener, colorant,preservative, thickening stabilizer, antioxidant, color former, bleach,fungicide, gum base, bittering agent, enzyme, gloss agent, acidulant,seasoning, emulsifier, enhancement agent, agent for production, flavor,spice, and the like.

When the nutritional composition is provided as food and drink, it canbe packaged as a single serving. Single serving packaging can be usedwhen the amount of food and drink to be ingested per meal is determinedin advance. Examples thereof include a single serving package such aspack, bag, bottle, box in case of drinks, gum, jelly, yogurt, cookie andthe like. The single serving package can be a pack, bag, and the like,in case of foods in the form of granule, powder, slurry, and the like.Particularly, when the food or drink are specified for health,nutritional, special-use, or invalid uses, the composition can bepackaged as a single serving unit amount, such as when the compositionis to be suspended or dissolved in a bottle to give a drink etc., for asingle consumption and the like.

The amount of the nutritional composition to be ingested per day can beindividually determined depending on the age, sex, body weight, mealcondition, and the like, and can be about 50 kcal-2000 kcal for an adultper day. This amount can be ingested in about 1 to 3 portions a day.When the nutritional composition is formulated in a single serving foodor drink in a package form of one ingestion amount unit, the amount tobe ingested one time as determined above can be individually packed.

The nutritional composition can be produced by conventional formulationtechniques and food production techniques.

The nutritional composition can be useful as a pharmaceutical product,food or drink, and the like for increasing muscle mass and/orsuppressing a decrease in muscle mass. In addition, the nutritionalcomposition can be useful as a pharmaceutical product, food or drink,and the like for increasing energy consumption. Furthermore, thenutritional composition can be useful as a pharmaceutical product, foodor drink, and the like for the prophylaxis and/or improvement of adecrease in muscle mass unaccompanied by an increase in inflammatorycytokine.

The nutritional composition can be useful as a pharmaceutical product,food and drink, and the like for the prevention and/or improvement ofmuscle weakness symptom. The muscle weakness symptom is also calledsarcopenia, decreases the amount of activity due to the decreasedmuscular strength of four limbs, and further causes chronic obstructivepulmonary diseases (COPD). The nutritional composition can effectivelyprevent and/or improve such muscle weakness symptom.

The nutritional composition can be useful as a pharmaceutical product,food or drink, and the like for enhancing rehabilitation effect formuscle recovery. Recovery of muscular strength by rehabilitation takestime, but the muscle function needs to be recovered before progressionof contracture. The composition can increase muscle mass and is usefulfor enhancing rehabilitation effect. It is also useful for theimprovement of low nutrient condition, improvement of deterioration inlocomotorium, prophylaxis and/or improvement of locomotive syndrome,prevention of falling and the like, as well as increase of muscle massin sports and the like.

The nutritional composition can be useful as a pharmaceutical product,food or drink, and the like for decreasing visceral fat. Increase invisceral fat induces insulin resistance, which can result in thedevelopment or progression of various diseases associated with insulinresistance. The nutritional composition can effectively reduce visceralfat.

The nutritional composition is useful as a pharmaceutical product, foodor drink, and the like for the prophylaxis and/or improvement ofdyslipidemia associated with visceral fat increase. When the visceralfat increases, dyslipidemia such as high LDL-cholesterolemia,hypertriglyceridemia, remnant hyperlipoproteinemia, high small denseLDL, and low HDL-cholesterolemia can occur. As a result of the onset ofdyslipidemia, the production of lipoperoxides such as LDL cholesterolperoxide increases, and the risk of developing atherosclerosis becomeshigh. The nutritional composition can effectively suppress the onset ofdyslipidemia such as increased blood LDL cholesterol associated withvisceral fat increase, and is useful for the prevention and/orimprovement of dyslipidemia associated with visceral fat increase.

The nutritional composition is useful as a pharmaceutical product, foodor drink, and the like for the prophylaxis and/or improvement ofhyperglycemia associated with visceral fat increase. Insulin resistancedue to visceral fat increase depresses glucose uptake efficiency fromblood into the cell, and causes hyperglycemia. The nutritionalcomposition is useful for the prophylaxis and/or improvement ofhyperglycemia associated with visceral fat increase.

The nutritional composition is useful as a pharmaceutical product, foodor drink, and the like for the prophylaxis and/or improvement of fattyliver and deterioration of liver function associated with visceral fatincrease. Mesenteric fat, which is representative of increased visceralfat, releases free fatty acids into the portal blood, and causesaccumulation of fat in the liver, which is the nearest organ. On theother hand, hyperinsulinemia due to insulin resistance also activatessynthesis of fat from sugar in the liver. Furthermore, fat accumulationin the liver aggravates liver function. The nutritional composition isuseful for the prophylaxis and/or improvement of fatty liver anddeterioration of liver function associated with visceral fat increase.

The nutritional composition can be useful as a pharmaceutical product,food or drink, and the like for the prophylaxis and/or improvement of adecrease in basal energy consumption. A decrease in the basal energyconsumption in elderly people means depressed energy utilization in thebody, which exerts a vast influence on the metabolism such as synthesisof proteins to be utilized as energy. In addition, when the energyutilization is depressed, the ingested energy becomes redundant, and theredundant energy accumulates as fat. The nutritional composition can beuseful for the prophylaxis and/or improvement of a decrease in the basalenergy consumption.

The nutritional composition can be useful as a pharmaceutical product,food or drink, and the like for the prophylaxis and/or improvement of adecrease in basal body temperature. When the body temperature is low,the metabolic activity becomes weak, and energy production and bodyprotein synthesis are depressed. Particularly, in elderly people sincethermoregulatory competence is also attenuated, a low body temperaturecannot be easily increased. The nutritional composition is useful forthe prophylaxis and/or improvement of a decrease in the basal bodytemperature.

The nutritional composition can be a multipurpose type comprehensivenutritional composition containing lysine and n-3 fatty acid, andfurther, one or more kinds of branched chain amino acids such as valine,leucine and/or isoleucine, in a powder form and the like.

The nutritional composition can be used for increasing muscle massand/or suppressing a further decrease in muscle mass of a person with adecreased muscle mass (e.g., elderly people, bedridden person etc.). Itis particularly effective when visceral fat associated withdyslipidemia, hyperglycemia, fatty liver, and/or deterioration of liverfunction is particularly used as an energy source for muscle proteinsynthesis. The nutritional composition greatly contributes to thosepeople having no effective nutritional composition heretofore.

EXAMPLES

The present invention is now explained based on the followingnon-limiting Examples.

Example 1 Preparation of Nutritional Composition

The nutritional composition shown in Table 1 was prepared with the goalof providing an optimum nutritional effect for elderly people. Theamounts of the starting materials of the liquid are shown in Table 2.Each component and an emulsifier were added to water, mixed, and anemulsion step was repeatedly performed in a high-pressure emulsifyingmachine under pressurization of 500-1,000 kg/cm² to give an emulsioncomposition. As the amino acid composition, L-leucine (0.38 g), L-valine(0.19 g), L-isoleucine (0.23 g) and L-lysine (0.32 g) were added per 100kcal of the composition. In this case, the total amount of the branchedchain amino acids (leucine, valine, isoleucine) including those derivedfrom a protein was 1.46 g per 100 kcal of the composition. The emulsioncomposition was placed in an aluminum bag by a conventional fillingmachine, and sterilized by a retort sterilization machine under generalconditions. The nutritional composition of this Example stably containedall ingredients one year later, and the viscosity at 25° C. was 9 mPa·s.A thickener was added to the regulated liquid, whereby a nutritionalcomposition with regulated thickness (1,000-7,000 mPa·s) or jelly couldbe produced. Furthermore, even when the amount of the amino acid addedwas 2-fold, similar production was possible. The calorie density of thenutritional composition was 1.0 kcal/ml.

TABLE 1 Example 1 nutritional composition component unit content/100kcal Protein source g 4.5 L-leucine g 0.38 L-valine g 0.19 L-isoleucineg 0.23 L-lysine g 0.32 lipid g 2.80 medium chain fatty acid g 0.54containing EPA mg 67 containing n-3 fatty acid g 0.195 hydrocarbonate g14.2 carbohydrates g 13 dietary fiber g 1.2 sodium mg 185 calcium mg 65iron mg 0.8 phosphorus mg 55 magnesium mg 26 potassium mg 130 copper mg0.11 iodine μg 13 manganese mg 0.34 selenium μg 2.5 zinc mg 1.65 chromeμg 2.5 molybdenum μg 2.1 vitamin A μg RE *1 81 vitamin D μg 0.46 vitaminE mg α-TE *2 2.73 vitamin K μg 6.3 vitamin B₁ mg 0.6 vitamin B₂ mg 0.36niacin mg NE *3 1.3 vitamin B₆ mg 0.6 folic acid μg 20 vitamin B₁₂ μg0.5 biotin μg 4.2 pantothenic acid mg 0.5 vitamin C mg 40.0 *1: retinolequivalents *2: amount based on α-tocopherol *3: niacin equivalents

TABLE 2 Example 1 starting material composition ◯ Example 1 compositionmixing ratio amount starting material (w/v %) added unit casein sodium3.8000 114 kg Lysine hydrochloride 0.4000 12.0 kg leucine 0.3833 11.5 kgisoleucine 0.2300 6.90 kg valine 0.1900 5.70 kg dextrin 11.07 332 kgsugar 2.367 71.0 kg edible vegetable oil 1.387 41.6 kg medium-chaintriglyceride 0.5367 16.1 kg fish oil 0.2633 7.90 kg citric acid 0.15934.78 kg trisodium citrate 0.5433 16.3 kg potassium carbonate 0.2280 6.84kg magnesium chloride 0.2207 6.62 kg calcium lactate 0.2960 8.88 kgglycerolcalcium phosphate 0.1303 3.91 kg mineral yeast premix *1 0.062931.888 kg Indigestible dextrin 1.5070 45.2 kg emulsifier 0.6606 19.8 kgβ-carotene oil 0.0015 45.0 g vitamin E 0.0052 156 g vitamin premix *20.1035 3.105 kg sodium ascorbate 0.0810 2.43 kg prepared water 83.082492 kg total 107.7 3231 kg starting material amount unit *1 mineralyeast premix composition molybdenum-containing yeast 13.0 gwater-soluble chrome-containing yeast 21.0 g selenium-containing yeast37.0 g manganese-containing yeast 203 g copper-containing yeast 337 gzinc-containing yeast 707 g biotin-containing yeast 344 g sodium ferrouscitrate 226 g total 1.888 kg *2 vitamin premix composition folic acid0.60 g vitamin D powder 8.50 g vitamin B 12 0.1% powder 15.0 griboflavin 5′-phosphate sodium 19.1 g calcium pantothenate 22.3 gpyridoxine hydrochloride 28.6 g nicotinic acid amide 38.5 g thiaminehydrochloride 40.7 g vitamin A powder 45.7 g vitamin K2 powder 58.0 gkelp extract powder 278 g sodium erythorbate 2550 g total 3.105 kg

Example 2 Preparation of Integrated Nutritional Composition

In the same manner as in Example 1, the nutritional composition shown inTable 3 was prepared. In the same manner as in Example 1, each componentand an emulsifier were added to water, mixed, and an emulsion step wasrepeatedly performed several times in a high-pressure emulsifyingmachine under pressurization of 50-500 kg/cm² to give an emulsioncomposition. As the amino acid composition, L-leucine (0.28 g), L-valine(0.14 g), L-isoleucine (0.14 g) and L-lysine (0.28 g) were added per 100kcal of the composition. In this case, the total amount of the branchedchain amino acids (leucine, valine, isoleucine) including those derivedfrom a protein is 1.30 g per 100 kcal of the composition. The emulsioncomposition was placed in an aluminum bag by a conventional fillingmachine, and sterilized by a retort sterilization machine under generalconditions. The nutritional composition of this Example stably containedall ingredients one year later, and the viscosity at 25° C. was 4,000mPa·s. An appropriate amount of a thickener was added to the regulatedliquid to further impart viscosity, whereby a nutritional compositionwith regulated thickness (1,000-7,000 mPa·s) or jelly could be produced.The calorie density of the nutritional composition was 2.0 kcal/ml.

TABLE 3 Example 2 nutritional composition component unit content/100kcal Protein source g 4.5 L-leucine g 0.28 L-valine g 0.14 L-isoleucineg 0.14 L-lysine g 0.28 lipid g 2.80 medium chain fatty acid g 0.60containing EPA mg 67 containing n-3 fatty acid g 0.19 hydrocarbonate g14.2 carbohydrates g 13 dietary fiber g 1.2 sodium mg 185 calcium mg 65iron mg 0.8 phosphorus mg 55 magnesium mg 26 potassium mg 130 copper mg0.11 iodine μg 13 manganese mg 0.34 selenium μg 2.5 zinc mg 1.65 chromeμg 2.5 molybdenum μg 2.1 vitamin A μg RE *1 81 vitamin D μg 0.46 vitaminE mg α-TE *2 2.73 vitamin K μg 6.3 vitamin B₁ mg 0.6 vitamin B₂ mg 0.36niacin mg NE *3 1.3 vitamin B₆ mg 0.6 folic acid μg 20 vitamin B₁₂ μg0.5 biotin μg 4.2 pantothenic acid mg 0.5 vitamin C mg 40.0 *1: retinolequivalents *2: amount based on α-tocopherol *3: niacin equivalents

Example 3 Preparation of Powder Nutritional Composition

The powder type nutritional composition shown in Table 4 was prepared.The amounts of the starting materials of the liquid are shown in Table5. Each component and an emulsifier were added to an appropriate amountof water, mixed, and an emulsion step was repeatedly performed severaltimes in a high-pressure emulsifying machine under pressurization of2-50 kg/cm² to give an emulsion composition. This emulsion compositionwas spray dried to give a powder nutritional composition. As the aminoacid composition, L-leucine (3.9 g), L-valine (2.4 g), L-isoleucine (2.0g) and L-lysine (4.1 g) were added per 100 kcal of the composition. Thispowder nutritional composition was placed in an aluminum bag by aconventional filling machine. The powder nutritional composition of thisExample stably contained all ingredients one year later. Thisnutritional composition could be used even after dissolving in warmwater and mixing with a nutritional supplement.

TABLE 4 Example 3 nutritional composition component unit content/100kcal Protein source g 12.4 L-leucine g 3.9 L-valine g 2.4 L-isoleucine g2.0 L-lysine g 4.1 lipid g 4.8 containing EPA mg 812 containing n-3fatty acid g 1.34 hydrocarbonate g 1.6 vitamin A μg RE *1 413 vitamin Emg α-TE *2 19.2 vitamin B₁ mg 5.3 vitamin B₂ mg 3.2 vitamin B₆ mg 5.1vitamin C mg 392.9 *1: retinol equivalents *2: amount based onα-tocopherol

TABLE 5 Example 3 starting material composition name of startingmaterials mixing ratio w/v % L-leucine 18.65 L-isoleucine 11.30 L-valine9.475 L-lysine hydrochloride 24.64 fish oil 22.99 vitamin A oil 500,000IU/g 0.01230 thiamine hydrochloride 0.02571 riboflavin 5′-phosphatesodium 0.02037 Pyridoxine hydrochloride 0.03038 tocopherol 40% 0.2146 Lascorbic acid Na 2.146 emulsifier 10.487 100.0

Experimental Example 1 Study of Enrichment Effect of Amino Acid and n-3Fatty Acid in Visceral Fat Increase Animal Model

The effects of amino acid and n-3 fatty acid in the nutritionalcomposition to improve visceral fat increase and muscle mass decreasewas examined by the following experiment. To be specific, 10-week-oldmale C57BL/6J mice were fed with the same high-fat diet as in 1B groupshown in Table 6 to prepare visceral fat increase models. They weredivided into 3 groups (each group N=5-9), and fed an experimental dietof the composition shown in Table 6 for 2 weeks. 1B group wascontinuously fed a high-fat diet containing a large amount of beef fatrich in saturated fatty acid. 1C group was fed an experimental diet of astandard composition. 1D group was fed an experimental diet with anenriched composition, wherein the total amount of protein and fat wasnot changed, and 0.32 g, 0.8 g, 0.24 g of lysine, BCAA (3 kinds ofbranched chain amino acid consisting of leucine, valine, isoleucine) andfish oil, respectively, per 100 kcal of experimental diet, were used forsubstitution. In this case, 1D group was fed with 1.46 g of the totalamount of BCAA including those derived from protein, 0.24 g of the totalamount of n-3 fatty acid and 67 mg of eicosapentaenoic acid (EPA), eachper 100 kcal of the experimental diet. 1A group as a normal controlgroup was fed an experimental diet with a standard compositionthroughout the test period, without a high-fat diet. Two weeks afteringestion of experimental diets, they were fasted for 16 hr, and themesenteric fat weight and gastrocnemius muscle weight were measured. Theresults are shown in FIG. 1.

TABLE 6 experimental diet composition table (content per 100 kcal ofexperimental diet) fat (g) n-3 fatty Protein source (g) fish oil acid L-L- L- L- beef (g) amount hydro- group casein lysine valine leucineisoleucine soybean fat containing contained carbonate constitution (g)(g) (g) (g) (g) oil (g) (g) EPA (mg) (g) (g) 1A group 4.5 2.80 0.17 15.41B group 4.5 0.56 5.17 0.05 7.2 1C group 4.5 2.80 0.17 15.4 1D group3.38 0.32 0.2 0.4 0.2 2.56 0.24 0.24 15.4 67

The body weight of the 1B group fed a high-fat diet was 1.2 times thatof the 1A group fed a general diet. Thereafter, throughout theevaluation period when the experimental diet was given, the body weightof the 1D group was lower than that of the 1C group, even though thecalorie intake was the same (1D: 31.7±3.3 g, 1C, 34.4±1.7 g). Moreover,as shown in FIG. 1, the mesenteric fat weight significantly increased inthe 1B group as compared to the 1A group, and the gastrocnemius muscleweight was significantly decreased in the 1B group as compared to the 1Agroup. On the other hand, the mesenteric fat weight was significantlydecreased in the 1D group and the gastrocnemius muscle weight wassignificantly increased, as compared to the 1C group. In this high-fatdiet ingestion model, CRP in plasma (C reactive protein) showed nodifference between groups (1A group: 7.7±2.4 ng/ml, 1B group: 9.0±3.3ng/ml, 1C group: 7.0±5.3 ng/ml, 1D group: 8.6±3.9 ng/ml).

From these test results, a diet enriched with lysine, BCAA,eicosapentaenoic acid can decrease mesenteric fat weight, which is theweight of visceral fat, and increase gastrocnemius muscle weight.

Experimental Example 2 Single Effect and Combined Effect of Amino Acids(Lysine, BCAA) and n-3 Fatty Acid

As a result of Experimental Example 1, enrichment with amino acid andn-3 fatty acid was shown to be necessary for decreasing mesenteric fatweight and increasing gastrocnemius muscle weight. However, it is notclear whether each component shows this effect alone. Therefore, aminoacid and n-3 fatty acid were tested either singly or in combination.That is, visceral fat increase models were prepared in the same manneras in Experimental Example 1, divided into 4 groups (each group N=5-9),and fed an experimental diet of the composition shown in Table 7 for 2weeks. The 2A group was fed an experimental diet of a standardcomposition. The 2D group was fed with an experimental diet an enrichedcomposition, wherein the total amount of protein and fat was notchanged, and 0.32 g, 0.8 g, 0.24 g of lysine, BCAA and fish oil,respectively, per 100 kcal of experimental diet, were used. In thiscase, the 2D group was fed 1.46 g of the total amount of BCAA includingthose derived from protein, 0.24 g of the total amount of n-3 fattyacid, and 67 mg of eicosapentaenoic acid (EPA), each per 100 kcal of theexperimental diet. Furthermore, the 2B group and 2C group were fed anexperimental diet enriched with amino acid or n-3 fatty acid. Two weeksafter ingestion of the experimental diet, they were fasted for 16 hr,and the mesenteric fat weight and gastrocnemius muscle weight weremeasured. The results are shown in FIG. 2.

TABLE 7 experimental diet composition table (content per 100 kcal ofexperimental diet) fat (g) fish oil Protein source (g) (g) casein L-lysL-val L-leu L-ile soybean with EPA n3 fatty acid hydro- (g) (g) (g) (g)(g) oil (g) (mg) amount (g) carbonate (g) 2A 4.5 2.80 0.17 15.4 2B 3.380.32 0.2 0.4 0.2 2.80 0.17 15.4 2C 4.5 2.56 0.24 0.24 15.4 67 2D 3.380.32 0.2 0.4 0.2 2.56 0.24 0.24 15.4 67

The total calorie intake was not different between groups duringingestion of the experimental diet. As shown in FIG. 2, the 2D groupsignificantly decreased mesenteric fat weight, which is the visceralfat, and significantly increased gastrocnemius muscle weight. Incontrast, decrease of mesenteric fat weight and increase ofgastrocnemius muscle weight in the 2B group and the 2C group wereclearly weak as compared to the effects in the 2D group, and the effectin the 2B group even combined with the effect in the 2C group was stilllower than the effect in the 2D group.

From these results, the mesenteric fat decreasing action andgastrocnemius muscle increasing action by the enriched diet was notprovided by the amino acid nor the n-3 fatty acid alone, but by asynergistic effect of the amino acid and n-3 fatty acid in combination.That is, in this test, the combined use of lysine and n-3 fatty acid,and further, branched chain amino acid was shown to be useful.

Experimental Example 3 Energy Metabolism Enhancing Effect andMitochondria Increasing Effect of Amino Acids and n-3 Fatty Acid

Then, the influence of the nutritional composition enriched with aminoacid and n-3 fatty acid on energy metabolism using an exhaled gasmetabolism measuring apparatus was examined. That is, visceral fatincrease model prepared in the same manner as in Experimental Example 1was fed an experimental diet with a standard composition (3A group) oran experimental diet with an enriched composition (3B group) shown inTable 8, and subjected to an evaluation. To stabilize the amount ofintake, a one-week experimental diet acclimation period was set, afterwhich one animal was placed in each cage within 2 weeks, and oxygenconsumption (ml/min) and carbon dioxide production (ml/min) weremeasured (metabolism measuring system for small animals MK-5000 RQ,Muromachi Kikai Co., Ltd.). During the measurement, an experimental dietwas given, and the amount of intake was measured. The respiratoryquotient was calculated by carbon dioxide production÷oxygen consumption,and the energy consumption was calculated by the formula of Weir [energyconsumption (kcal/min)=3.9 (kcal)×oxygen consumption (ml/min)+1.1(kcal)×carbon dioxide production (ml/min)]. The results are shown inFIG. 3.

TABLE 8 experimental diet composition table (content per 100 kcal ofexperimental diet) fat (g) fish oil Protein source (g) (g) casein L-lysL-val L-leu L-ile soybean with EPA n3 fatty acid hydro- (g) (g) (g) (g)(g) oil (g) (mg) amount (g) carbonate (g) 3A 4.5 2.80 0.17 15.4 3B 3.380.32 0.2 0.4 0.2 2.56 0.24 0.24 15.4 67

As shown in FIG. 3, the respiratory quotient of the 3B group decreasedsignificantly as compared to the 3A group during the diet ingestionperiod (9:00-16:00; diet) and the dark period after eating (16:00-19:00;dark period), and also decreased significantly as compared to the 3Agroup even in the light period (19:00-9:00: light period), which is freeof a direct influence of eating. The energy consumption of the 3B groupincreases significantly or increases as compared to the 3A group at anyperiod.

In addition, the ability of the nutritional composition to effect themitochondria was examined. That is, visceral fat increase model preparedin the same manner as in Experimental Example 1 was fed an experimentaldiet with a standard composition (3A group) or an experimental diet withan enriched composition (3B group) shown in Table 8, and, on the lastday of the test, the liver and gastrocnemius muscle were rapidlycollected and cryopreserved. DNA was extracted from each of thecryopreserved liver (100 mg) and gastrocnemius muscle (200 mg) by usingISOGEN (Japan gene). To 50 ng DNA were added the primer of d-loop ofmitochondria shown in Table 9 and SYBER GREEN Master Mix (ABI), andamplification and quantification analysis were performed using an ABI7700 detector. For the amendment of DNA amount, gene cyclophillin A(intron code part) on the chromosome was used. As for the synthesizedprimer, the occurrence of desired production was confirmed by agaroseelectrophoresis after PCR reaction.

TABLE 9 DNA primer of mitochondria and chromosome Gene sense antisensech ACACGCCATAATGGCACTGG CAGTCTTGGCAGTGCAGAT cyclo- (Sequence Listing(Sequence Listing phillin SEQ ID NO: 1) SEQ ID NO: 2) A mt d-CGCAAAACCCAATCACCTAA TTGGGGTTTGGCATTAAGAG loop (Sequence Listing(Sequence Listing SEQ ID NO: 3) SEQ ID NO: 4)

Since mitochondria plays an important role in fat combustion in thecell, the number of intracellular mitochondria was analyzed. While onlyone intranuclear chromosomal DNA is present in the cell, the number ofmitochondria is not the same. Therefore, the amount of mitochondria inthe cell was assumed by a method of determining the ratio of the DNAamount of mitochondrial coding gene and that of the nuclear coding gene.The results are shown in FIG. 4.

As shown in FIG. 4, the mitochondrial DNA amount of the 3B groupsignificantly increased in both the liver and gastrocnemius muscle, ascompared to the 3A group.

By this test, a nutritional composition containing lysine and n-3 fattyacid, and further, a branched chain amino acid, can result in anincrease in the number of mitochondria to enhance fat utilization, andcan increase energy consumption that leads to an increase in the bodytemperature, and the like.

INDUSTRIAL APPLICABILITY

The nutritional composition as described herein is effective forincreasing muscle mass and/or suppressing a decrease in muscle mass. Inaddition, the nutritional composition is effective for increasing energyconsumption. In addition, the nutritional composition is particularlyuseful for decreasing visceral fat by increasing energy consumption byenhanced fat utilization and the like. In addition, it is useful for theprophylaxis and/or improvement of dyslipidemia, hyperglycemia, fattyliver, and deterioration of liver function associated with visceral fatincrease. Furthermore, by increasing muscle mass or suppressing adecrease in muscle mass by fat energy supply and the like, thenutritional composition is also useful for the prophylaxis and/orimprovement of sarcopenia, chronic obstructive pulmonary disease (COPD),promotion of rehabilitation effect for muscle recovery, improvement oflow nutrient condition, improvement of deterioration in locomotorium,prophylaxis and/or improvement of locomotive syndrome, prevention offalling, increasing muscle mass in sports and the like. Moreover, thenutritional composition is effective for the prophylaxis and/orimprovement of a decrease in basal energy consumption, and prophylaxisand/or improvement of a decrease in basal body temperature. Furthermore,the nutritional composition is highly safe, and can be used for a longtime without placing an excessive protein load even for elderly peoplewith attenuated kidney function.

While the invention has been described in detail with reference topreferred embodiments thereof, it will be apparent to one skilled in theart that various changes can be made, and equivalents employed, withoutdeparting from the scope of the invention. Each of the aforementioneddocuments is incorporated by references herein in its entirety.

1. A nutritional composition comprising: i) n-3 fatty acid, and ii) aningredient selected from the group consisting of free lysine, dipeptidescontaining lysine and lysine salts, and combinations thereof; whereinthe ingredient is present in the composition in an amount of 0.1 g-10.0g per 100 kcal of the composition, and wherein the n-3 fatty acid ispresent in the composition in an amount of 0.17 g-5.00 g per 100 kcal ofthe composition.
 2. The nutritional composition according to claim 1,further comprising a branched chain amino acid selected from the groupconsisting of valine, leucine, isoleucine, and combinations thereof. 3.The nutritional composition according to claim 2, wherein the branchedchain amino acid is present in the composition in an amount of 1.0 g-20g per 100 kcal of the composition.
 4. The nutritional compositionaccording to claim 2, wherein the branched chain amino acid in a freeform is present in the composition in an amount of 0.1 g-15 g per 100kcal of the composition.
 5. The nutritional composition according toclaim 1, wherein the n-3 fatty acid is selected from the groupconsisting of eicosapentaenoic acid, docosapentaenoic acid,docosahexaenoic acid, and combinations thereof.
 6. The nutritionalcomposition according to claim 5, comprising 10 mg-3000 mg ofeicosapentaenoic acid per 100 kcal of the composition.
 7. Thenutritional composition according to claim 1, further comprising avitamin selected from the group consisting of: 0.2 mg-20.0 mg of vitaminB1 per 100 kcal of the composition, 0.25 mg-20.0 mg of vitamin B2 per100 kcal of the composition, 0.3 mg-20.0 mg of vitamin B6 per 100 kcalof the composition, 0.3 μg-20.0 μg of vitamin B12 per 100 kcal of thecomposition, and combinations thereof.
 8. The nutritional compositionaccording to claim 1, further comprising a vitamin selected from thegroup consisting of: 50 μg retinol equivalents-2500 μg retinolequivalents of vitamin A per 100 kcal of the composition, 10 mg-700 mgof vitamin C per 100 kcal of the composition, 1 mg-50 mg of vitamin Ebased on α-tocopherol per 100 kcal of the composition, and combinationsthereof.
 9. The nutritional composition according to claim 1, furthercomprising 1 mg-50 mg of zinc per 100 kcal of the composition and/or 1.0μg-50.0 μg of selenium per 100 kcal of the composition.
 10. Thenutritional composition according to claim 1, comprising medium chainfatty acid oil in an amount of 10 wt %-65 wt % relative to the totalamount of lipid.
 11. The nutritional composition according to claim 1,which is capable of increasing muscle mass and/or suppressing a decreasein the muscle mass.
 12. The nutritional composition according to claim1, which is capable of decreasing visceral fat or increasing energyconsumption.
 13. The nutritional composition according to claim 1, whichis capable of an effect selected from the group consisting of i)prophylaxis and/or improvement of muscle weakness symptom, ii) promotionof rehabilitation effect for muscle recovery, iii) prophylaxis and/orimprovement of dyslipidemia associated with visceral fat increase, iv)prophylaxis and/or improvement of hyperglycemia associated with visceralfat increase, v) prophylaxis and/or improvement of fatty liverassociated with visceral fat increase, vi) prophylaxis and/orimprovement of deteriorated liver function associated with visceral fatincrease, vii) prophylaxis and/or improvement of decrease in basalenergy consumption, viii) prophylaxis and/or improvement of decrease inbasal body temperature, ix) prophylaxis and/or improvement of musclemass decrease unaccompanied by an increase in inflammatory cytokine, x)improvement of low nutrient condition, xi) improvement of deteriorationin locomotorium, xii) prophylaxis and/or improvement of locomotivesyndrome, xiii) prevention of falling, ixv) increase of muscle mass insports, and xv) combinations thereof.
 14. A method for increasing musclemass and/or suppressing a decrease in muscle mass, comprisingadministering an effective amount of the composition according toclaim
 1. 15. A method for decreasing visceral fat or increasing energyconsumption, comprising administering an effective amount of thecomposition according to claim
 1. 16. A method for treating a conditioncomprising administering an effective amount of the compositionaccording to claim 1, wherein said condition is selected from the groupconsisting of i) the prophylaxis and/or improvement of muscle weaknesssymptom, ii) the promotion of rehabilitation effect for muscle recovery,iii) the prophylaxis and/or improvement of dyslipidemia associated withvisceral fat increase, iv) the prophylaxis and/or improvement ofhyperglycemia associated with visceral fat increase, v) the prophylaxisand/or improvement of fatty liver associated with visceral fat increase,vi) the prophylaxis and/or improvement of deteriorated liver functionassociated with visceral fat increase, vii) the prophylaxis and/orimprovement of decrease of basal energy consumption, viii) theprophylaxis and/or improvement of decrease of basal body temperature,ix) the prophylaxis and/or improvement of muscle mass decreaseunaccompanied by an increase in inflammatory cytokine, x) theimprovement of low nutrient condition, xi) the improvement ofdeterioration in locomotorium, xii) the prophylaxis and/or improvementof locomotive syndrome, xiii) prevention of falling, ixv) a method forincreasing muscle mass in sports, and xv) combinations thereof.
 17. Amethod for producing an agent for increasing muscle mass and/orsuppressing a decrease in muscle mass comprising formulating acomposition according to claim
 1. 18. A method for producing an agentfor decreasing visceral fat or increasing energy consumption comprisingformulating a composition according to claim
 1. 19. A method forproducing an agent for treating a condition comprising formulating acomposition according to claim 1, wherein the condition is selected fromthe group consisting of: i) the prophylaxis and/or improvement of muscleweakness symptom, ii) promoting a rehabilitation effect for musclerecovery, iii) the prophylaxis and/or improvement of dyslipidemiaassociated with visceral fat increase, iv) the prophylaxis and/orimprovement of hyperglycemia associated with visceral fat increase, v)the prophylaxis and/or improvement of fatty liver associated withvisceral fat increase, vi) the prophylaxis and/or improvement ofdeteriorated liver function associated with visceral fat increase, vii)the prophylaxis and/or improvement of decrease of basal energyconsumption, viii) the prophylaxis and/or improvement of decrease ofbasal body temperature, ix) the prophylaxis and/or improvement of musclemass decrease unaccompanied by an increase in inflammatory cytokine, x)improvement of low nutrient condition, xi) improving deterioration inlocomotorium, xii) the prophylaxis and/or improvement of locomotivesyndrome, ixv) prevention of falling, xv) increasing muscle mass insports, and xvi) combinations thereof.